Abstract
Purinergic signalling in rat GFSHR-17 granulosa cells was characterised by Ca2+-imaging and perforated patch-clamp. We observed a resting intracellular Ca2+-concentration ([Ca2+]i) of 100 nM and a membrane potential of −40 mV. This was consistent with high K+− and Cl− permeability and a high intracellular Cl− concentration of 40 mM. Application of ATP for 5–15 s every 3 min induced repeated [Ca2+]i increases and a 30 mV hyperpolarization. The phospholipase C inhibitor U73122 or the IP3-receptor antagonist 2-aminoethoethyl diphenyl borate suppressed ATP responses. Further biochemical and pharmacological experiments revealed that ATP responses were related to stimulation of P2Y2 and P2Y4 receptors and that the [Ca2+]i increase was a prerequisite for hyperpolarization. Inhibitors of Ca2+-activated channels or K+ channels did not affect the ATP-evoked responses. Conversely, inhibitors of Cl− channels hyperpolarized cells to −70 mV and suppressed further ATP-evoked hyperpolarization. We propose that P2Y2 and P2Y4 receptors in granulosa cells modulate Cl− permeability by regulating Ca2+-release.
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Bintig, W., Baumgart, J., Walter, W.J. et al. Purinergic signalling in rat GFSHR-17 granulosa cells: an in vitro model of granulosa cells in maturing follicles. J Bioenerg Biomembr 41, 85–94 (2009). https://doi.org/10.1007/s10863-009-9199-5
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DOI: https://doi.org/10.1007/s10863-009-9199-5